Biochemical features of the wild type and the evolved UPO variant
| Biochemical or spectroscopy featureh | Value | |
|---|---|---|
| wtUPO1e | PaDa-I mutantf | |
| Mass (Da)a | 46,000 | 52,000 |
| Mass (Da)b | NDg | 51,100 |
| Mass (Da)c | 35,942 | 35,914 |
| Glycosylation degree (%) | 22 | 30 |
| Thermal stability (T50) (°C)d | 53 | 55 |
| pI | 4.9–5.7 | 5.5 |
| Optimum pH for ABTS | 4.0 | 4.0 |
| Optimum pH for DMP | 7.0 | 6.0 |
| Optimum pH for NBD | 6.5 | 6.0 |
| Rz (A418/A280) | 2.4 | 1.8 |
| Soret region (nm) | 420 | 418 |
| CT1 (nm) | 572 | 570 |
| CT2 (nm) | 540 | 537 |
↵a Estimated by SDS-PAGE.
↵b Estimated by MALDI-TOF mass spectrometry.
↵c Estimated from the amino acid composition.
↵d Estimated from purified variants.
↵e wtUPO1, UPO1 wild type expressed in A. aegerita.
↵f PaDa-I mutant, the ultimate variant of the whole evolution process in S. cerevisiae (containing the evolved signal peptide [n*] plus the evolved UPO1) (see Fig. S1 in the supplemental material).
↵g ND, not determined.
↵h CT1 and CT2, charge transference bands 1 and 2, respectively.